RESTORING THE RARE KENTUCKY LADY’S RESTORING THE RARE KENTUCKY LADY’S SLIPPER ORCHID IN THE KISATCHIE NATIONAL SLIPPER ORCHID IN THE KISATCHIE NATIONAL

FOREST: OUTPLANTING STUDY RESULTSFOREST: OUTPLANTING STUDY RESULTS

Jim Barnett, Emeritus ScientistJim Barnett, Emeritus ScientistU.S. Forest ServiceU.S. Forest ServiceSouthern Research Station, andSouthern Research Station, andSt d C di tSt d C di tStudy Coordinator, Study Coordinator, Central LA Orchid SocietyCentral LA Orchid Society

David Moore BotanistDavid Moore BotanistDavid Moore, BotanistDavid Moore, BotanistU.S. Forest ServiceU.S. Forest ServiceKisatchie National ForestKisatchie National Forest

Kevin Allen, Science Teacher Kevin Allen, Science Teacher Captain Shreve High SchoolCaptain Shreve High SchoolShreveport, LAShreveport, LAp ,p ,

Cypripedium kentuckienseCypripedium kentuckiense

CHARACTERISTICS OF KENTUCKY LADY’S CHARACTERISTICS OF KENTUCKY LADY’S SLIPPER (SLIPPER (Cypripedium kentuckiense Cypripedium kentuckiense C.F. ReedC.F. Reed))

A tall, stately perennial plant A tall, stately perennial plant with large flowerswith large flowers

Underground rhizomes allowUnderground rhizomes allow Underground rhizomes allow Underground rhizomes allow periods of plant dormancyperiods of plant dormancy

Its range is the SE United Its range is the SE United States, but it is rare States, but it is rare throughout the rangethroughout the range

Kisatchie National Forest hasKisatchie National Forest hasKisatchie National Forest has Kisatchie National Forest has 4 sites on 600,000 acres.4 sites on 600,000 acres.

Occurs in mesic forests on Occurs in mesic forests on t l id tt l id tstream slopes or acid steep stream slopes or acid steep

sitessites

An Opportunity…An Opportunity…An Opportunity…An Opportunity… Kevin Allen, amateur Kevin Allen, amateur

b t i tb t i t d HS t d td HS t d tbotanist botanist and HS student and HS student in 2004in 2004——hand pollinated hand pollinated orchidsorchidsorchids.orchids.

After 3 years of After 3 years of monitoring, discovered amonitoring, discovered amonitoring, discovered a monitoring, discovered a seedpod at a Catahoula seedpod at a Catahoula Ranger District site.Ranger District site.

Kevin has completing his Kevin has completing his master’s degree and master’s degree and teaching in a Shreveportteaching in a Shreveportteaching in a Shreveport teaching in a Shreveport high school.high school.

It’s Alive…It’s Alive… Kevin sent his seed Kevin sent his seed

pod to Spangle Creekpod to Spangle Creekpod to Spangle Creek pod to Spangle Creek Lab for germination.Lab for germination.

Seedpod proved toSeedpod proved toSeedpod proved to Seedpod proved to contain 1000s of contain 1000s of viable seeds!viable seeds!

Kevin contacted the Kevin contacted the KNF with his idea of KNF with his idea of rere--introduction.introduction.

A COOPERATIVE EFFORTA COOPERATIVE EFFORTA COOPERATIVE EFFORTA COOPERATIVE EFFORT The KNF contacted the Central Louisiana The KNF contacted the Central Louisiana

Orchid Society (CLOS) and Southern Research Orchid Society (CLOS) and Southern Research y ( )y ( )Station (SRS) on a native orchid reintroduction Station (SRS) on a native orchid reintroduction project.project.

CLOS was receptive. A research proposal was CLOS was receptive. A research proposal was developed to fund the project.developed to fund the project.

The Southwest Regional Orchid Grower’s The Southwest Regional Orchid Grower’s Association (SWROGA) provided a grant to buy Association (SWROGA) provided a grant to buy plants and the project began in 2006.plants and the project began in 2006.

Additional grants from the Forest Service were Additional grants from the Forest Service were obtained in 2007 and 2008.obtained in 2007 and 2008.

OBJECTIVES OF THE STUDYOBJECTIVES OF THE STUDYOBJECTIVES OF THE STUDYOBJECTIVES OF THE STUDY Primarily to reintroduce up to several hundred rare Primarily to reintroduce up to several hundred rare

hid t KNF l dhid t KNF l dorchids to KNF lands.orchids to KNF lands. Research various growing media and horticultural Research various growing media and horticultural

fftechniques from seedling to juvenile plant.techniques from seedling to juvenile plant. Reintroduction methods: site selection, season, Reintroduction methods: site selection, season,

protection, age.protection, age. In the spring of 2006 & 2007 plantlets were In the spring of 2006 & 2007 plantlets were

distributed to 10 orchid growers for development distributed to 10 orchid growers for development of cultural techniques.of cultural techniques.

Growers kept records of their techniques.Growers kept records of their techniques.

EARLY SEEDLING EARLY SEEDLING DEVELOPMENTDEVELOPMENT

EARLY SEEDLING EARLY SEEDLING DEVELOPMENTDEVELOPMENTDEVELOPMENTDEVELOPMENTDEVELOPMENTDEVELOPMENT

Early seedling growth Early seedling growth was rapid. These was rapid. These seedlings were about 4seedlings were about 4seedlings were about 4 seedlings were about 4 weeks old.weeks old.

Two or three leavesTwo or three leavesTwo or three leaves Two or three leaves developed quickly.developed quickly.

Most of the seedlings Most of the seedlings ggthat were received that were received were vigorous and were vigorous and healthyhealthyhealthy.healthy.

Many growers used trays like

thisthis.Based on 2006 results, the medium used in 2007the medium used in 2007 was a commercial mix (Promix) with addition of(Promix) with addition of sand. This combination resulted in a loose mix that favored seedling development and lifting f l tifor planting.

Dormancy “recognized”Dormancy “recognized”Dormancy “recognized”Dormancy “recognized”Dormancy recognizedDormancy recognizedDormancy recognizedDormancy recognized

Photos on October 9, 2006 of seedlings with dead and live shoots. Root systems of these seedlings were washed to evaluate condition.

Photos on October 9, 2006 of seedlings with dead and live shoots. Root systems of these seedlings were washed to evaluate condition.

Status of root systems of Status of root systems of dli d d li ?dli d d li ?

Status of root systems of Status of root systems of dli d d li ?dli d d li ?seedlings dormant and alive?seedlings dormant and alive?seedlings dormant and alive?seedlings dormant and alive?

Condition of root systems of seedlings with a recently dead shoot (left) and one that has a green shoot (right). Both root systems are live and the seedling on the left is in a dormant state.

Condition of root systems of seedlings with a recently dead shoot (left) and one that has a green shoot (right). Both root systems are live and the seedling on the left is in a dormant state.

Size of planting stockSize of planting stock Bill Cullina, New England Wildflower Society, Bill Cullina, New England Wildflower Society,

indicates that 1indicates that 1--yearyear--old seedlings have been old seedlings have been yy ggoutplanted with success. However, older plants outplanted with success. However, older plants may result in better establishment.may result in better establishment.

Lifting seedlings from traysLifting seedlings from traysg g yg g y Different methods were tested Different methods were tested

to evaluate the best way to liftto evaluate the best way to liftto evaluate the best way to lift to evaluate the best way to lift seedlings from the trays.seedlings from the trays.

The method that worked best The method that worked best was to lift a block of the was to lift a block of the medium and a seedling with a medium and a seedling with a spatulaspatulaspatula.spatula.

Since the media is loose, the Since the media is loose, the seedlings are easily separated seedlings are easily separated without injury.without injury.

Most seedling still have the Most seedling still have the dead shoot growthdead shoot growthdead shoot growth.dead shoot growth.

ROOT DEVELOPMENT OF 2006 AND ROOT DEVELOPMENT OF 2006 AND 2007 SEEDLINGS AT PLANTING2007 SEEDLINGS AT PLANTING

Before planting numbers ofBefore planting numbers of 2006 S dli Before planting, numbers of Before planting, numbers of roots and growing points were roots and growing points were counted on each plant.counted on each plant.

2006 Seedlings

Averages of data followAverages of data follow::2006 Crop 2007 Crop2006 Crop 2007 Crop

2007 Seedlings

Rep.Rep. Roots Roots (#)(#)

Shoots Shoots (#)(#)

Roots Roots (#)(#)

Shoots Shoots (#)(#)

11 7.67.6 1.01.0 9.09.0 1.21.22007 Seedlings

2007 Seedlings22 6.26.2 1.21.2 6.66.6 1.41.4

33 6.66.6 1.01.0 7.27.2 1.01.0

44 5.65.6 1.01.0 10.210.2 1.61.644 5.65.6 1.01.0 10.210.2 1.61.6

Avg.Avg. 6.56.5 1.01.0 8.38.3 1.31.3

NATIVE CYPRIPEDIUM SEEDLINGS ON NATIVE CYPRIPEDIUM SEEDLINGS ON THE CATAHOULA RANGER DISTRICTTHE CATAHOULA RANGER DISTRICTTHE CATAHOULA RANGER DISTRICTTHE CATAHOULA RANGER DISTRICT

T o nati e C pripedi m seedlingsT o nati e C pripedi m seedlingsTwo native Cypripedium seedlings Two native Cypripedium seedlings located on the Kisatchie National located on the Kisatchie National

Forest’s Catahoula Ranger District.Forest’s Catahoula Ranger District. Photo taken April 15, 2008Photo taken April 15, 2008

PLANTING SITES/SOILSPLANTING SITES/SOILS

Kentucky lady’s slippers Kentucky lady’s slippers require a site with few require a site with few hours of direct sun a day. hours of direct sun a day. and filtered or dappledand filtered or dappledand filtered or dappled and filtered or dappled shade for the rest.shade for the rest.S il h ld b i tS il h ld b i t Soils should be moist, Soils should be moist, but not wet, and but not wet, and reasonably well drainedreasonably well drainedreasonably well drained.reasonably well drained.

The pH should be slightly The pH should be slightly acid to nearly neutralacid to nearly neutralacid to nearly neutral. acid to nearly neutral.

INDICATOR PLANTSINDICATOR PLANTS

Plants found on typical lady’s Plants found on typical lady’s slipper sites are beech and hollyslipper sites are beech and hollyslipper sites are beech and holly slipper sites are beech and holly trees, Christmas ferns, and trees, Christmas ferns, and cranefly orchids.cranefly orchids.yy

Tipularia discolorTipularia discolor is a is a frequent orchid on the frequent orchid on the

it It lit It lsites. Its leaves are sites. Its leaves are bright green with a bright green with a purple back in winter. purple back in winter. p pp pThe leaves die when The leaves die when flower stalks appear in flower stalks appear in the summerthe summerthe summer.the summer.

Layout of outplanting test, four Layout of outplanting test, four replications of five seedlingsreplications of five seedlingsreplications of five seedlingsreplications of five seedlings

PF07* PS08 PS08 PF072006 2006

PS08 PF07

S082007

02006

PF07 PS08PS082007

PF072007

PF072007

PS082006

*In each 5 seedling plot the top identification e g PF07

Rep 1 Rep 2

*In each 5-seedling plot, the top identification, e.g., PF07, represents the season of outplanting and the lower date represents the year of the crop.

PLANTING TECHNIQUESPLANTING TECHNIQUES Work compost into the soil.Work compost into the soil.

Spread roots to preserve the Spread roots to preserve the p pp proot pattern and dribble soil to root pattern and dribble soil to cover.cover.

When the plant is almost When the plant is almost covered, keep tamping the soil covered, keep tamping the soil lightly as you fill while holding lightly as you fill while holding the bud(s) so as to keep them the bud(s) so as to keep them

t d tht d that a proper depth.at a proper depth.

When complete the bud should When complete the bud should be just breaking the surfacebe just breaking the surface..

Cover the site with mulch.Cover the site with mulch.

Seedling characteristics and survival of various Seedling characteristics and survival of various treatments when planted on December 18, 2007 and treatments when planted on December 18, 2007 and

March 6, 2008March 6, 2008Characteristics at plantingCharacteristics at planting

S i lS i lPlanting variablePlanting variable

SurvivalSurvival# Roots# Roots # Shoots# Shoots % 4/1/08% 4/1/08 % 7/7/08% 7/7/08

D b 18 2007 l tiD b 18 2007 l tiDecember 18, 2007 plantingDecember 18, 2007 planting2006 crop2006 crop 6.5 6.5 aa 1.01.0 7070 25252007 crop2007 crop 8.38.3 1.31.3 5050 1515pp

March 6, 2008 plantingMarch 6, 2008 planting2007 unfertilized2007 unfertilized 8.48.4 1.31.3 8585 1515

a a The numbers represent an average of 4 replications of 5 seedlings each.The numbers represent an average of 4 replications of 5 seedlings each.

2007 fertilized 2007 fertilized bb 8.18.1 1.21.2 6565 1010

p g p gp g p gb b SlowSlow--release nutrients (one teaspoon of Osmocote 19release nutrients (one teaspoon of Osmocote 19--66--12) were applied at each 12) were applied at each planting spot after each seedling was planted.planting spot after each seedling was planted.

Treatments imposed to seedlings at the Treatments imposed to seedlings at the December 30 2008 planting of 2007 stockDecember 30 2008 planting of 2007 stockDecember 30, 2008 planting of 2007 stock.December 30, 2008 planting of 2007 stock.

Depth of planting is theDepth of planting is theDepth of planting is the Depth of planting is the primary treatment being primary treatment being evaluated in this testevaluated in this test——1/2 1/2

d 1 i h d thd 1 i h d thand 1 inch depth.and 1 inch depth. Seedling size/age Seedling size/age

comparisons will be madecomparisons will be madecomparisons will be made comparisons will be made with earlier plantings.with earlier plantings.

In a related test, In a related test, i l ti ith f li l ti ith f linoculation with fungal inoculation with fungal material is being evaluated material is being evaluated in side test.in side test.

Performance of 2007 crop seedlings grown in trays, lifted, Performance of 2007 crop seedlings grown in trays, lifted, and planted December 30, 2008, and measured in the field and planted December 30, 2008, and measured in the field

M 20 2009 d A il 6 2010M 20 2009 d A il 6 2010May 20, 2009 and April 6, 2010May 20, 2009 and April 6, 2010

Variables measuredVariables measuredPlanting Treatments Planting Treatments аа

Sh llSh ll DDVariables measuredVariables measured ShallowShallow DeepDeep

Characteristics at plantingCharacteristics at planting——December 30, 2008December 30, 2008

Roots (number)Roots (number) 10 810 8 bb 10 210 2Roots (number)Roots (number) 10.8 10.8 bb 10.210.2

Shoots (number)Shoots (number) 1.01.0 1.01.0

Measurements on May 20 2009Measurements on May 20 2009Measurements on May 20, 2009Measurements on May 20, 2009

Survival (percent)Survival (percent) 9292 6060

Measurements on April 6, 2010Measurements on April 6, 2010

aa The planting depths were ½ inch (shallow) and 1 inch (deep) below the The planting depths were ½ inch (shallow) and 1 inch (deep) below the

Measurements on April 6, 2010Measurements on April 6, 2010

Survival (percent)Survival (percent) 1515 1010

p g p ( ) ( p)p g p ( ) ( p)soil surfacesoil surface.bb The numbers presented are averages of 40 seedlings (8 reps of 5 The numbers presented are averages of 40 seedlings (8 reps of 5 seedlings each).seedlings each).

CONCLUSIONSCONCLUSIONS Shallow planting is more successful Shallow planting is more successful

than deepthan deep Fertilizing at planting was not Fertilizing at planting was not

successfulsuccessful If dormant when planted season ofIf dormant when planted season of Stage of development is Stage of development is If dormant when planted, season of If dormant when planted, season of

planting is not importantplanting is not important Seedling age or stage of Seedling age or stage of Effects of fungal Effects of fungal

inoculationinoculation

g pg pimportant for successimportant for success

development is very importantdevelopment is very important Caging of planting site is not Caging of planting site is not

neededneeded

inoculation inoculation of potted seedlingsof potted seedlings

SeedlingSeedlingtreatmenttreatment

Survival at Survival at 6 months6 monthsneededneeded

Inoculation with mycorrhizae seems Inoculation with mycorrhizae seems importantimportant

treatmenttreatment 6 months6 monthsInoculatedInoculatedwith soilwith soilhumushumus

52 percent52 percent

Additional research is needed to Additional research is needed to establish reintroduction guidelinesestablish reintroduction guidelines

Control Control (not(not

inoculated)inoculated)

37 percent37 percent

CONTINUING RESEARCHCONTINUING RESEARCH Grants have been received to fund Grants have been received to fund

Kevin Allen’s high school honors Kevin Allen’s high school honors chemistry class’ efforts to developchemistry class’ efforts to developchemistry class efforts to develop chemistry class efforts to develop CypripediumCypripedium seed germination and seed germination and developmental techniques.developmental techniques.Ke in Allen ill train ith Dr Zettler atKe in Allen ill train ith Dr Zettler at Kevin Allen will train with Dr. Zettler at Kevin Allen will train with Dr. Zettler at Illinois College to begin fungal relation Illinois College to begin fungal relation studies with seed germination and studies with seed germination and seedling establishmentseedling establishmentseedling establishment.seedling establishment.

Plantlets from these germination Plantlets from these germination studies will in grown in garden studies will in grown in garden situations to develop larger plants forsituations to develop larger plants forsituations to develop larger plants for situations to develop larger plants for field outplanting. field outplanting.

Tests of seedling size at planting, Tests of seedling size at planting, depth of planting fungal inoculationdepth of planting fungal inoculation

Our goal is to Our goal is to reintroduce these reintroduce these

depth of planting, fungal inoculation, depth of planting, fungal inoculation, and planting site will continue.and planting site will continue.

orchidsorchids